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Impaired angiogenesis and tumor development by inhibition of the mitotic kinesin Eg5.

Exertier P, Javerzat S, Wang B, Franco M, Herbert J, Platonova N, Winandy M, Pujol N, Nivelles O, Ormenese S, Godard V, Becker J, Bicknell R, Pineau R, Wilting J, Bikfalvi A, Hagedorn M - Oncotarget (2013)

Bottom Line: Chemical blockade of Eg5 inhibits endothelial cell proliferation and migration in vitro.Mitosis-independent vascular outgrowth in aortic ring cultures is strongly impaired after Eg5 or Mklp2 protein inhibition.In vivo, interfering with KIF11/Eg5 function causes developmental and vascular defects in zebrafish and chick embryos and potent inhibition of tumor angiogenesis in experimental tumor models.

View Article: PubMed Central - PubMed

Affiliation: University Bordeaux, LAMC, UMR 1029, F-33405 Talence, France.

ABSTRACT
Kinesin motor proteins exert essential cellular functions in all eukaryotes. They control mitosis, migration and intracellular transport through interaction with microtubules. Small molecule inhibitors of the mitotic kinesin KiF11/Eg5 are a promising new class of anti-neoplastic agents currently evaluated in clinical cancer trials for solid tumors and hematological malignancies. Here we report induction of Eg5 and four other mitotic kinesins including KIF20A/Mklp2 upon stimulation of in vivo angiogenesis with vascular endothelial growth factor-A (VEGF-A). Expression analyses indicate up-regulation of several kinesin-encoding genes predominantly in lymphoblasts and endothelial cells. Chemical blockade of Eg5 inhibits endothelial cell proliferation and migration in vitro. Mitosis-independent vascular outgrowth in aortic ring cultures is strongly impaired after Eg5 or Mklp2 protein inhibition. In vivo, interfering with KIF11/Eg5 function causes developmental and vascular defects in zebrafish and chick embryos and potent inhibition of tumor angiogenesis in experimental tumor models. Besides blocking tumor cell proliferation, impairing endothelial function is a novel mechanism of action of kinesin inhibitors.

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Effects of kinesin blockade in chick and zebrafish embryos(a) Control chick embryos showed normal expansion of the allantoic vesicle (al) at day 4.5 (HH24). Arrows point to the border of the vesicle. (b, c) Eg5 inhibition leads to complete arrest of CAM development; only a rudimentary tissue with a primitive vascular network develops. (d) Blockade of Eg5 function using ispinesib (ISP) from 24 to 48 hpf leads to a significant increase of embryos with modification of the posterior blood islands (PBI) or reduction of blood cells at 1 and 3μM. (e) Phenotype-score of Tg(kdrl:EGFP)s843 embryos injected with indicated morpholinos (Mo) at 48 hpf. At doses higher than 0.5 ng, most embryos die and show severe edema and circulation defects. (e, f) At 0.2 ng, 46% of embryos displayed a mild phenotype with a curved and shortened tail and normal circulation, 34% had severe circulation defects, including pericardial edema (arrow), and 8% of embryos were dead (70× magnification). (g) Fluorescence micrographs of control and kif11 morphants (115× magnification). Asterisk and arrows denote random vascular defects. DLAV = dorsal longitudinal anastomotic vessel, ISV = intersomitic vessels.
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Figure 5: Effects of kinesin blockade in chick and zebrafish embryos(a) Control chick embryos showed normal expansion of the allantoic vesicle (al) at day 4.5 (HH24). Arrows point to the border of the vesicle. (b, c) Eg5 inhibition leads to complete arrest of CAM development; only a rudimentary tissue with a primitive vascular network develops. (d) Blockade of Eg5 function using ispinesib (ISP) from 24 to 48 hpf leads to a significant increase of embryos with modification of the posterior blood islands (PBI) or reduction of blood cells at 1 and 3μM. (e) Phenotype-score of Tg(kdrl:EGFP)s843 embryos injected with indicated morpholinos (Mo) at 48 hpf. At doses higher than 0.5 ng, most embryos die and show severe edema and circulation defects. (e, f) At 0.2 ng, 46% of embryos displayed a mild phenotype with a curved and shortened tail and normal circulation, 34% had severe circulation defects, including pericardial edema (arrow), and 8% of embryos were dead (70× magnification). (g) Fluorescence micrographs of control and kif11 morphants (115× magnification). Asterisk and arrows denote random vascular defects. DLAV = dorsal longitudinal anastomotic vessel, ISV = intersomitic vessels.

Mentions: DMN injection into the allantoic vesicle at Hamburger&Hamilton stage 21 (HH21) embryos completely inhibited expansion of the vesicle (HH24; Fig 5b, c). Only a rudimentary tissue mass with a primitive vascular network developed. This effect was consistent in all DMN-injected embryos, control embryos showed normal growth of the allantoic vesicle (arrows, Fig. 5a).


Impaired angiogenesis and tumor development by inhibition of the mitotic kinesin Eg5.

Exertier P, Javerzat S, Wang B, Franco M, Herbert J, Platonova N, Winandy M, Pujol N, Nivelles O, Ormenese S, Godard V, Becker J, Bicknell R, Pineau R, Wilting J, Bikfalvi A, Hagedorn M - Oncotarget (2013)

Effects of kinesin blockade in chick and zebrafish embryos(a) Control chick embryos showed normal expansion of the allantoic vesicle (al) at day 4.5 (HH24). Arrows point to the border of the vesicle. (b, c) Eg5 inhibition leads to complete arrest of CAM development; only a rudimentary tissue with a primitive vascular network develops. (d) Blockade of Eg5 function using ispinesib (ISP) from 24 to 48 hpf leads to a significant increase of embryos with modification of the posterior blood islands (PBI) or reduction of blood cells at 1 and 3μM. (e) Phenotype-score of Tg(kdrl:EGFP)s843 embryos injected with indicated morpholinos (Mo) at 48 hpf. At doses higher than 0.5 ng, most embryos die and show severe edema and circulation defects. (e, f) At 0.2 ng, 46% of embryos displayed a mild phenotype with a curved and shortened tail and normal circulation, 34% had severe circulation defects, including pericardial edema (arrow), and 8% of embryos were dead (70× magnification). (g) Fluorescence micrographs of control and kif11 morphants (115× magnification). Asterisk and arrows denote random vascular defects. DLAV = dorsal longitudinal anastomotic vessel, ISV = intersomitic vessels.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3926828&req=5

Figure 5: Effects of kinesin blockade in chick and zebrafish embryos(a) Control chick embryos showed normal expansion of the allantoic vesicle (al) at day 4.5 (HH24). Arrows point to the border of the vesicle. (b, c) Eg5 inhibition leads to complete arrest of CAM development; only a rudimentary tissue with a primitive vascular network develops. (d) Blockade of Eg5 function using ispinesib (ISP) from 24 to 48 hpf leads to a significant increase of embryos with modification of the posterior blood islands (PBI) or reduction of blood cells at 1 and 3μM. (e) Phenotype-score of Tg(kdrl:EGFP)s843 embryos injected with indicated morpholinos (Mo) at 48 hpf. At doses higher than 0.5 ng, most embryos die and show severe edema and circulation defects. (e, f) At 0.2 ng, 46% of embryos displayed a mild phenotype with a curved and shortened tail and normal circulation, 34% had severe circulation defects, including pericardial edema (arrow), and 8% of embryos were dead (70× magnification). (g) Fluorescence micrographs of control and kif11 morphants (115× magnification). Asterisk and arrows denote random vascular defects. DLAV = dorsal longitudinal anastomotic vessel, ISV = intersomitic vessels.
Mentions: DMN injection into the allantoic vesicle at Hamburger&Hamilton stage 21 (HH21) embryos completely inhibited expansion of the vesicle (HH24; Fig 5b, c). Only a rudimentary tissue mass with a primitive vascular network developed. This effect was consistent in all DMN-injected embryos, control embryos showed normal growth of the allantoic vesicle (arrows, Fig. 5a).

Bottom Line: Chemical blockade of Eg5 inhibits endothelial cell proliferation and migration in vitro.Mitosis-independent vascular outgrowth in aortic ring cultures is strongly impaired after Eg5 or Mklp2 protein inhibition.In vivo, interfering with KIF11/Eg5 function causes developmental and vascular defects in zebrafish and chick embryos and potent inhibition of tumor angiogenesis in experimental tumor models.

View Article: PubMed Central - PubMed

Affiliation: University Bordeaux, LAMC, UMR 1029, F-33405 Talence, France.

ABSTRACT
Kinesin motor proteins exert essential cellular functions in all eukaryotes. They control mitosis, migration and intracellular transport through interaction with microtubules. Small molecule inhibitors of the mitotic kinesin KiF11/Eg5 are a promising new class of anti-neoplastic agents currently evaluated in clinical cancer trials for solid tumors and hematological malignancies. Here we report induction of Eg5 and four other mitotic kinesins including KIF20A/Mklp2 upon stimulation of in vivo angiogenesis with vascular endothelial growth factor-A (VEGF-A). Expression analyses indicate up-regulation of several kinesin-encoding genes predominantly in lymphoblasts and endothelial cells. Chemical blockade of Eg5 inhibits endothelial cell proliferation and migration in vitro. Mitosis-independent vascular outgrowth in aortic ring cultures is strongly impaired after Eg5 or Mklp2 protein inhibition. In vivo, interfering with KIF11/Eg5 function causes developmental and vascular defects in zebrafish and chick embryos and potent inhibition of tumor angiogenesis in experimental tumor models. Besides blocking tumor cell proliferation, impairing endothelial function is a novel mechanism of action of kinesin inhibitors.

Show MeSH
Related in: MedlinePlus